Target (protein/gene name): Dihydropteroate synthase 2 E.C Number: 2.5.1.15 NCBI Gene # or RefSeq#: NC_000962.3 PBD ID: 2VP8 Organism: Mycobacterium Tuberculosis Etiologic Risk Group: N/A Disease Information: Mycobacterium is an organism that induces the disease Tuberculosis. Mycobacterium does this through the formation of aerosol droplets onto lung-alveolar spaces in the body. The disease was it's first sighting in dirt found on earth but was soon found in animal, specifically cows, which in turn passed on what was called M.bovis (M.tuberculosis) to humans. It was found that over the years the disease has been quite fatal and it wasn't until the 20th century that vaccines and antibiotics were discovered to treat this disease. Although there are discovered treatments for this disease, there is still fatal cases around the world today. The most commons symptoms seen affect the lungs and nervous system.
Essentiality: Dihydropteroate synthase participates in the biosynthetic pathways for folate (in bacteria, plants and fungi) and methanopterin (in archaea). The enzyme exists in varying types of multifunctional proteins in different organisms such as the plant Arabidopsis thaliana and the yeast Saccharomyces cerevisiae.
A simple method to detect plasmodium falciparum single nucleotide polymorphisms in many proteins including Dihydropteroate synthase using polymerase chain reaction and enzyme linked immunosorbent assay-based technology.
Query Coverage: 100% Max % Identities: 100% % Positives: 100% Chain used for Homology: A
Similarity to Human: No similarity to human was found
Inhibitors: In bacteria, antibacterial sulfonamides act as competitive inhibitors of the enzyme dihydropteroate synthase, DHPS. DHPS catalyses the conversion of PABA (para-aminobenzoate) to dihydropteroate, a key step in folate synthesis.
Expression: The genes for dihydropteroate synthase of Mycobacterium tuberculosis were expressed under control of the T5 promoter in a dihydropteroate synthase-deficient strain of Escherichia coli. Using three chromatography steps, M. tuberculosis can be purified.
Do Not Need this info for Spring (but still copy these lines to your Target page for now) Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): (link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol) -- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
E.C Number: 2.5.1.15
NCBI Gene # or RefSeq#: NC_000962.3
PBD ID: 2VP8
Organism: Mycobacterium Tuberculosis
Etiologic Risk Group: N/A
Disease Information: Mycobacterium is an organism that induces the disease Tuberculosis. Mycobacterium does this through the formation of aerosol droplets onto lung-alveolar spaces in the body. The disease was it's first sighting in dirt found on earth but was soon found in animal, specifically cows, which in turn passed on what was called M.bovis (M.tuberculosis) to humans. It was found that over the years the disease has been quite fatal and it wasn't until the 20th century that vaccines and antibiotics were discovered to treat this disease. Although there are discovered treatments for this disease, there is still fatal cases around the world today. The most commons symptoms seen affect the lungs and nervous system.
Links:
1. What is Tuberculosis?
2. Mycobacterium tuberculosis Pathogenesis and Molecular Determinants of Virulence
Essentiality:
Dihydropteroate synthase participates in the biosynthetic pathways for folate (in bacteria, plants and fungi) and methanopterin (in archaea). The enzyme exists in varying types of multifunctional proteins in different organisms such as the plant Arabidopsis thaliana and the yeast Saccharomyces cerevisiae.
Link:
Expasy Page
Druggable Target:
A Promising Target for Novel Anti-Malarial Chemotherapy
Assayable Enzyme Target:
http://www.ajtmh.org/docserver/fulltext/14761645/72/2/0720155.pdf?expires=1525621478&id=id&accname=guest&checksum=7B23BE897925EAA8A03DB545CD85E784
A simple method to detect plasmodium falciparum single nucleotide polymorphisms in many proteins including Dihydropteroate synthase using polymerase chain reaction and enzyme linked immunosorbent assay-based technology.
Brenda:
http://www.brenda-enzymes.org/enzyme.php?ecno=2.5.1.15
Structure:
Pairwise alignment of BlastP:
<span style="background-color: #ffffff; color: #222222; font-size: 11.869px;">Query 1 MNKSLIIFGIVNITSDSFSDGGRYLAPDAAIAQARKLMAEGADVIDLGPASSNPDAAPVS 60 MNKSLIIFGIVNITSDSFSDGGRYLAPDAAIAQARKLMAEGADVIDLGPASSNPDAAPVS Sbjct 13 MNKSLIIFGIVNITSDSFSDGGRYLAPDAAIAQARKLMAEGADVIDLGPASSNPDAAPVS 72 Query 61 SDTEIARIAPVLDALKADGIPVSLDSYQPATQAYALSRGVAYLNDIRGFPDAAFYPQLAK 120 SDTEIARIAPVLDALKADGIPVSLDSYQPATQAYALSRGVAYLNDIRGFPDAAFYPQLAK Sbjct 73 SDTEIARIAPVLDALKADGIPVSLDSYQPATQAYALSRGVAYLNDIRGFPDAAFYPQLAK 132 Query 121 SSAKLVVMHSVQDGQADRREAPAGDIMDHIAAFFDARIAALTGAGIKRNRLVLDPGMGFF 180 SSAKLVVMHSVQDGQADRREAPAGDIMDHIAAFFDARIAALTGAGIKRNRLVLDPGMGFF Sbjct 133 SSAKLVVMHSVQDGQADRREAPAGDIMDHIAAFFDARIAALTGAGIKRNRLVLDPGMGFF 192 Query 181 LGAAPETSLSVLARFDELRLRFDLPVLLSVSRKSFLRALTGRGPGDVGAATLAAELAAAA 240 LGAAPETSLSVLARFDELRLRFDLPVLLSVSRKSFLRALTGRGPGDVGAATLAAELAAAA Sbjct 193 LGAAPETSLSVLARFDELRLRFDLPVLLSVSRKSFLRALTGRGPGDVGAATLAAELAAAA 252 Query 241 GGADFIRTHEPRPLRDGLAVLAALKETARIR 271 GGADFIRTHEPRPLRDGLAVLAALKETARIR Sbjct 253 GGADFIRTHEPRPLRDGLAVLAALKETARIR 283 </span>Query Coverage: 100%Max % Identities: 100%
% Positives: 100%
Chain used for Homology: A
Similarity to Human: No similarity to human was found
Inhibitors:
In bacteria, antibacterial sulfonamides act as competitive inhibitors of the enzyme dihydropteroate synthase, DHPS. DHPS catalyses the conversion of PABA (para-aminobenzoate) to dihydropteroate, a key step in folate synthesis.
Folate Inhibitors
Expression:
The genes for dihydropteroate synthase of Mycobacterium tuberculosis were expressed under control of the T5 promoter in a dihydropteroate synthase-deficient strain of Escherichia coli. Using three chromatography steps, M. tuberculosis can be purified.
http://citeseerx.ist.psu.edu/viewdoc/download?doi=10.1.1.577.1166&rep=rep1&type=pdf
Transmembrane Bound Regions:
There are 4 Transmembrane Bound regions
TMpred graph Image
TMpred page Link:
https://embnet.vital-it.ch/cgi-bin/TMPRED_form_parser
CDS Gene Sequence:
Protein Sequence:
Do Not Need this info for Spring (but still copy these lines to your Target page for now)
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):
(link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)
-- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.